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Microfibril-associated glycoproteins attenuating pulmonary fibrosis

微纤维相关糖蛋白减轻肺纤维化

基本信息

批准号：
10595656
负责人：
Jeffrey Koenitzer
金额：
$ 15.62万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-04-01 至 2027-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10595656
关键词：
Acceleration Advisory Committees Affinity Attenuated Binding Bioinformatics Biological Assay Biology Bleomycin Bone Marrow Cell Nucleus Cells Characteristics Chromatin Collaborations Compensation Data Deposition Development Disease Disease Progression Exhibits Extracellular Matrix Family member Fibroblasts Fibrosis Follow-Up Studies Gene Expression Gene Expression Regulation Genes Glycoproteins Goals Growth Factor Homeostasis Human Hypoxia Image Immune Immune response In Situ Individual Inflammatory Inflammatory Response Pathway Injury Interstitial Lung Diseases Knock-out Knockout Mice Lung Lung Transplantation Lung diseases MFAP1 gene Macrophage Mediating Mentors Mesenchymal Microfibrils Modeling Mus Names Pathogenesis Pathway interactions Phase Phenotype Physicians Play Population Process Production Proliferating Protein Secretion Proteins Pulmonary Fibrosis Recovery Regulation Reporting Research Personnel Role Scientist Signal Transduction Specificity Supportive care Tissues Training Transforming Growth Factor beta Transposase Up-Regulation Work airway obstruction antifibrotic treatment career career development cytokine epigenome epigenomics experimental study fibrogenesis fibrotic lung idiopathic pulmonary fibrosis immune cell infiltrate immune function improved in vivo inhibitor interest lung histology lung injury mortality novel strategies receptor response side effect single cell sequencing single nucleus RNA-sequencing single-cell RNA sequencing transcriptome transcriptome sequencing transcriptomics transforming growth factor beta3

项目摘要

Project Summary/Abstract Idiopathic pulmonary fibrosis (IPF) is a progressive lung disease with limited treatment options and unclear pathogenesis. Extracellular matrix (ECM) deposition by proliferating fibroblast populations drives the progressive airflow limitation and hypoxia characteristic of IPF, mediated by the cytokine transforming growth factor beta (TGFB). Direct inhibition of TGFB leads to intolerable side effects, and there is increased interest in therapies that instead modulate its regulation. Microfibril-associated glycoproteins 1 and 2 (Magp1 and Magp2, or Magps) are fibroblast-produced proteins which are secreted to ECM, anchor to microfibrils, and bind TGFB to limit its signaling under basal conditions, but have no known role in fibrosis. Magp knockout (KO) mice exhibit increased TGFB signaling in various tissues and have immune cell alterations due to bone marrow TGFB effects. In a single cell RNASeq screen, the genes encoding these proteins were markedly upregulated in fibroblasts in fibrotic lungs, and combined KO of Magp1 and 2 led to more severe fibrosis after bleomycin injury in mice. This suggests that Magps act as inhibitors of fibrotic signaling and follow up studies in Magp1 and 2 individual KO mice show that deficiency of both proteins is required to produce a pro-fibrotic phenotype. Which fibroblasts populations produce Magps and how they limit fibrosis is unknown. We hypothesize that Magp expression is an adaptive response to lung injury and limits fibrosis by sequestering TGFB in ECM and will explore this central hypothesis through three specific aims: 1) Evaluate anti-fibrotic contributions of Magp1 and 2 in pulmonary fibrosis. 2) Evaluate Magp effects on TGFB signaling and immune responses in lung fibrosis. 3) Characterize Magp-producing fibroblast populations. Using combinations of KO and inducible/tissue-specific KO mice, we will determine the individual relationships of these proteins to fibrosis and whether they exhibit redundant antifibrotic functions in vivo. We will analyze alterations in TGFB signaling and immune cell infiltration in lung as possible underlying mechanisms of Magp protection from pulmonary fibrosis. Finally, we will employ our recently developed single nucleus RNASeq approach along with single nucleus assay for transposase-accessible chromatin (snATAC)-Seq to characterize the fibroblast populations that express Magp1 and 2 and evaluate how lung-wide transcriptomes and epigenomes change in the absence of Magps during fibrosis. This proposal will provide Dr. Koenitzer with the mentored training in single cell sequencing and bioinformatics, fibroblast and ECM biology, and lung histology/imaging needed to achieve his goal of independence as a physician-scientist. Under the oversight of an expert scientific advisory committee, He will realize career milestones, complete formal coursework, and develop lasting collaborations. Findings from this work will have implications beyond IPF in other interstitial lung diseases and enable Dr. Koenitzer to develop new approaches to the treatment of lung disease as an independent investigator.

项目总结/摘要特发性肺纤维化（IPF）是一种进行性肺部疾病，治疗选择有限且尚不清楚发病机制细胞外基质（ECM）沉积的增殖成纤维细胞群体驱动的， IPF的进行性气流受限和缺氧特征，由细胞因子转化生长介导因子β（TGFB）。TGFB的直接抑制导致不可耐受的副作用，并且对TGFB的直接抑制的兴趣增加。而不是调节其调节的疗法。微纤维相关糖蛋白1和2（Magp 1和Magp 2，或Magps）是成纤维细胞产生的蛋白质，其分泌到ECM，锚于微纤维，并结合TGFB 以限制其在基础条件下的信号传导，但在纤维化中没有已知的作用。Magp敲除（KO）小鼠在各种组织中表现出增加的TGFB信号传导，并且由于骨髓炎而具有免疫细胞改变。 TGFB效应。在单细胞RNASeq筛选中，编码这些蛋白质的基因显著上调，在纤维化肺的成纤维细胞中，Magp 1和2的联合KO导致博来霉素治疗后更严重的纤维化。小鼠损伤。这表明Magps可作为纤维化信号传导的抑制剂，并对Magp 1进行了后续研究。和2个单独的KO小鼠显示，两种蛋白质的缺陷是产生促纤维化表型所必需的。哪些成纤维细胞群体产生Magps以及它们如何限制纤维化尚不清楚。我们假设 Magp表达是对肺损伤的适应性反应，并通过在ECM中隔离TGFB来限制纤维化，将通过三个具体目标探索这一中心假设：1）评价抗纤维化的贡献肺纤维化中的Magp 1和Magp 2 2)评估Magp对TGFB信号传导和免疫的影响肺纤维化的反应。3)表征产生Magp的成纤维细胞群。使用组合 KO和诱导型/组织特异性KO小鼠，我们将确定这些蛋白质与纤维化以及它们是否在体内表现出多余的抗纤维化功能。我们将分析TGFB的改变肺中的信号传导和免疫细胞浸润可能是Magp保护肺纤维化最后，我们将使用我们最近开发的单核RNASeq方法，沿着转座酶可及染色质单核测定（snATAC）-Seq，用于表征成纤维细胞表达Magp 1和2的人群，并评估肺范围内的转录组和表观基因组在在纤维化过程中没有Magps。该提案将为Koenitzer博士提供以下方面的指导培训：单细胞测序和生物信息学，成纤维细胞和ECM生物学，以及肺组织学/成像，实现他作为一名医生兼科学家的独立目标。在专家科学顾问的监督下他将实现职业生涯的里程碑，完成正式的课程，并发展持久的合作。这项工作的发现将对IPF以外的其他间质性肺疾病产生影响，并使Dr。 Koenitzer作为独立研究者开发治疗肺部疾病的新方法。